Quantification of bindings of organometallic ruthenium complexes to GSTπ by mass spectrometry

Journal of Inorganic Biochemistry
Yu LinFuyi Wang

Abstract

Electrospray ionization mass spectrometry (ESI-MS) has been widely used to identify binding sites of metal complexes to proteins. However, the MS quantification of the metal-protein coordination remains a challenge. We have recently demonstrated by ESI-MS analysis that organometallic ruthenium complexes [(η(6)-arene)Ru(en)Cl](+) (arene=p-cymene (1), biphenyl (2) or 9,10-dihydrophenanthrene (3); en=ethylenediamine) bound to human glutathione-S-transferase π (GSTπ) at Cys15 and Cys48 within the G-site, and Cys102 and Met92 on the interface of the GSTπ dimer, showing inhibitory potency against the enzyme (J. Inorg. Biochem., 128 (2013) 77-84). Herein, we developed a mass spectrometric method to quantify the binding stoichiometry of the three complexes to GSTπ. The differences in signal intensities of the heavy-labelled peptides produced by tryptic digestion of the ruthenated GSTπ complexes and the respective light-labelled peptides in the tryptic digest of equimolar GSTπ were used to calculate the binding stoichiometry at specific residues. The results indicated that the pre-complexation of GSTπ with its substrate GSH significantly reduced the bindings of the ruthenium complexes at Met92 and Cys102, but had little impact on the bi...Continue Reading

References

May 10, 2002·Journal of Chromatography. a·Asish Chakraborty, Fred E Regnier
Oct 1, 2005·Chembiochem : a European Journal of Chemical Biology·Isam KhalailaPaul J Dyson
Oct 6, 2011·Proteomics·Maria StefanopoulouDirk A Wolters
Jan 17, 2012·The Journal of Biological Chemistry·Bengt Mannervik

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Citations

Sep 24, 2015·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Ileana DragutanAlbert Demonceau
Feb 1, 2019·Dalton Transactions : an International Journal of Inorganic Chemistry·Anamarija BrišJana Roithová
Feb 28, 2019·Rapid Communications in Mass Spectrometry : RCM·Yumiao HanFuyi Wang

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